KR102513930B1 - Anisoxanthos Flavidus Extract for Cosmetic Use - Google Patents

Abstract

The present invention relates in particular to the use of an anisoxanthos plavidus extract as an anti-aging agent to reduce the appearance of fine lines and wrinkles, to lift, smooth, tone, restructure or reshape the skin.

Description

Anisoxanthos Flavidus Extract for Cosmetic Use

The present invention relates to the field of cosmetics, in particular to anti-aging cosmetic agents.

The skin is the first barrier that protects an organism from external aggression. This organ is made up of several layers of tissue. It is divided into the outermost part of the skin, epidermis, dermis, connective tissue composed of fibroblasts and extracellular matrix and performing skin cohesive and nutritional functions, and subcutaneous tissue composed of adipocytes.

The epidermis is composed of several cell layers of keratinocytes. This includes, inter alia, the germ layer of the epidermis, called the basal layer, which contains dermal stem cells, the spiny layer ( Stratum spinosum ) consisting of several layers of polygonal cells, and one to three containing intracytoplasmic inclusion bodies. The granular layer ( stratum granulosum ) composed of canine layers of squamous cells, the keratohyalin granule and, finally, the keratin, called corneocyte, corresponding to the last stage of differentiation of keratinocytes. It is divided into horny layer ( stratum corneum ) composed of abundant non-nucleated cells.

The outermost cells of the stratum corneum are successively removed and replaced by cells of the lower layers in a process known as desquamation. Cell regeneration of the stratum corneum is based on a cell maturation process in which cells of the basal layer of the epidermis differentiate and gradually migrate through the various strata of the epidermis until they reach the stratum corneum in the form of keratinocytes.

Skin aging includes dysfunction of cellular differentiation and/or regenerated skin, which results in atrophy of all layers of the skin, whether due to normal phenomena of aging or enhanced by external factors such as exposure to ultraviolet radiation. From a histological point of view, inter alia, a decrease in the quality of the dermis is observed, in particular a decrease in the thickness of the epidermis and a loss of consistency of the extracellular matrix. Thus, it was observed that the collagen fibers in the extracellular matrix were fewer and less regular, and the ratio of type III collagen to type I collagen increased. The number of elastic fibers in the papillary dermis also decreases and the elastic fibers in the reticular dermis change. From an aesthetic point of view, these changes result in a modification of the skin's appearance and mechanical properties: it becomes less smooth, more rough, microreliefs become more pronounced, and over time microreliefs form deep wrinkles. line can be displayed. The skin also exhibits loss of elasticity, tone and firmness, which can lead to tissue sagging or sagging and tightening of wrinkles, particularly on the face or neck. Skin aging can affect the uniformity of the complexion and can lead to the appearance of pigment spots.

There are numerous anti-aging cosmetics on the market that attempt to prevent or attenuate skin sagging.

Nonetheless, there still remains a need for new anti-aging actives for preventing or treating the changes that currently result from skin aging, particularly those associated with skin sagging.

The present invention relates to cosmetic use of Anigozanthos flavidus extract as an anti-aging agent. In particular, the Anigoxanthos plavidus extract can be used as an agent for anti-wrinkle, smoothing, toning, restructuring, or remodeling for the skin.

Generally, the Anisoxanthos plavidus extract is present as an active agent in a cosmetic composition intended to prevent, retard and/or treat one or more signs of skin aging. Signs of skin aging include the appearance of fine lines and/or wrinkles in the skin, particularly on the lips or eyelids, withering or sagging of the skin, especially on the neck, loss of skin density, and loss of skin firmness. loss, loss of skin tonicity, loss of skin elasticity, loss of skin contractility, loss of skin maintenance, skin sagging, change in smooth appearance of skin, increase in skin roughness and change in facial contour It can be selected from the group consisting of.

Anisoxanthos plavidus extract can be obtained from aerial parts of Anisoxanthos plavidus , such as flowers, leaves, stems and combinations thereof. In some embodiments, the Anisoxanthos plavidus extract is obtained by extraction under conditions of a polar solvent, preferably high temperature.

As described above, the Anisoxanthos plavidus extract may be present as an active agent in a cosmetic composition. The cosmetic composition is preferably a vitamin, a sun filter and sunscreen, an anti-aging or wrinkle agent, an antioxidant, a lifting agent, a firming agent, an anti-spot agent, an anti-redness agent, and a slimming agent. slimming agent, dehydrating agent, humectant, emollient, scrubbing or exfoliating agent, matting agent, sebum control agent, lightening active, self-tanning active, It may include at least one additional cosmetic agent selected from the group consisting of tanning accelerators and combinations thereof.

In some embodiments, the cosmetic composition is an aqueous solution, an aqueous-alcoholic solution, an oil-in-water (O/W) or water-in-oil (W/O) or multiple (triple: W/O/W or O/W/O) emulsion, nano Emulsions, in particular O/W nanoemulsions with droplet sizes of less than 100 nm, aqueous gels or dispersions of the fatty phase in an aqueous phase with spherules, suspensions, preferably suspensions in aqueous or aqueous-alcoholic media , liposomal suspensions, powders, lotions, milks, creams, salves, gels, foams and ointments. Cosmetic compositions are cosmetics, make-up products or body hygiene products, such as lotions, milks, serums, aqueous or oily gels, emulsions, creams, in particular day or night creams, cream- gels, skin care waters, ointments, balms, foundations, sprays, eye shadows, sticks, lipsticks, glosses, lip balms, foams, deodorants, nourishing masks, shower gels, and exfoliating or scrubbing products.

The present invention also relates to a cosmetic composition for use in the prophylaxis or treatment of signs of aging, said composition comprising in particular an Anissanthos plavidus extract as defined above and one or more pharmaceutically acceptable excipients. According to a further aspect, the present invention is a "precursor" composition, i.e. a composition for the preparation of a cosmetic composition, the composition comprising:

- from 10% to 50%, preferably from 20% to 40% of the Anisoxanthos plavidus extract, preferably the Anisoxanthos plavidus liquid extract,

- 50% to 90%, preferably 60%, preferably selected from aqueous solvents, organic solvents, preferably ethanol, lower alcohols such as propanediol, butylene glycol, glycerol or isopropanol, lipophilic agents and mixtures thereof % to 80% carrier, and

- 0.1 optionally, preferably selected from pH regulators, antioxidants, preservatives, stabilizers, vectorizing agents, thickeners, emulsifiers, hydrophilic or lipophilic gelling agents, flavoring agents, mineral oils or organic oils and combinations thereof % to 30% of additional cosmetically acceptable excipients,

Percentages represent weight percentages relative to the total weight of the composition.

For example, the composition for preparing a cosmetic composition,

- 20% to 40% of an aqueous extract of Anisoxanthos plavidus , and

- contains 60% to 80% glycerol,

Percentages represent weight percentages relative to the total weight of the composition.

An object of the present invention is also a process for preparing a cosmetic composition according to the present invention, the process comprising adding a "precursor" composition according to the present invention to at least one acceptable excipient from a cosmetic point of view and/or at least one further active agent having a cosmetic effect. and mixing with

Figure 1 shows the amount of procollagen I, expressed as a percentage relative to the control, produced by fibroblasts cultured with TGF-β or with a composition at a concentration of 0.1%, referred to as "extract" (Example Example 1). An “extract” composition contains 5% by weight of an aqueous extract of Anisoxanthos plavidus flower, less than 35% by weight of water, and at least 50% by weight of glycerol.
Figure 2 is cultured with fetal calf serum (FCS), cultured with a 0.2% concentration "extract" composition (extract 0.2%), and the presence of a 0.1% concentration "extract" composition (extract 0.1%). Shows the amount of Tenascin-X (Tenascin-X) expressed as a percentage with respect to the control, produced by fibroblasts cultured under (Example 2).
3A-3C show the results regarding the effect of “extract” on the contractile force of human fibroblasts from the bottom of the wrinkle (BW) (Example 4). Figure 3A shows BW for 24 hours in the presence of 0.1% (v/v) of the "extract" composition, in the presence of 0.2% (v/v) of the "extract" composition, and in the absence of an active agent (control (-)). It represents the change in contractile force exerted by fibroblasts.
The bar graph in FIG. 3B corresponds to the area under the curve (AUC) of the curve in FIG. 3A. The bar graph in FIG. 3C corresponds to the maximum contractile force relative to the curve in FIG. 3A.
BW: BW fibroblasts without activator (negative control), BW fibroblasts cultured in the presence of an "extract" composition of activator 0.1%: 0.1% (v/v); and BW fibroblasts cultured in the presence of an "extract" composition of active agent 0.2%: 0.2% (v/v).
Figure 4 shows immunolabeling images of α-SM actin fibers in fibroblasts from the bottom of the wrinkles: (A): control; (B): after incubation with 0.2% of "extract"; and (C): after incubation with "extract" at 0.1%.
5A and 5B show the contractile force exerted by an acellular skin equivalent over time in the presence of an “extract” composition (active 0.2%) (Kangaroo Paw 0.2%) and in the absence of said composition (control). indicate
Figure 6 shows the change in the depth of crow's feet in faces treated with a placebo cream (Cream A) compared to a cream containing 2% of the "extract" (Cream B) (1 hour after application of the extract and 28 days after treatment). ).

Angioxanthos plavidus is an endemic plant species of the Haemodoraceae family, found primarily in southwestern Australia. This plant is characterized by flowers that are usually yellow or red and resemble kangaroo paws. To the Applicant's knowledge, Anisoxanthos plavidus extract has been used in cosmetics at best as a conditioning agent, for example in shampoo compositions. It appears that no cosmetic biological activity has been reported for Angioxanthos plavidus extract.

Applicants prepared an aqueous extract of Angioxanthos plavidus and studied the biological activity of this extract.

Surprisingly, the applicants have demonstrated that Xanthos plavidus extracts show activity against biological targets that play a key role in the organization and composition of the extracellular matrix of the skin. As exemplified in the examples, angioxanthos plavidus extract can stimulate the synthesis of type I procollagen by dermal fibroblasts and increase the synthesis of type XVII collagen by keratinocytes. Applicants also found that Angioxanthos plavidus extract can stimulate the synthesis of Tenascin-X, a glycoprotein that plays a key role in the structure and stability of elastic fibers and the organization of collagen fibers in the extracellular matrix of the skin. proved. Angioxanthos plavidus extract also stimulates the synthesis of actin alpha 2 (also known as alpha-smooth muscle actin (α-SMA)) in fibroblasts, resulting in the production of fibroblasts from the bottom of the wrinkles. increase the force of contraction. Angioxanthos plavidus extract has been found to be able to increase the contractile force exerted by its acellular skin equivalent, indicating an immediate contractile effect of the extract on the fibers of the dermis through physical action.

The applicant also conducted a clinical trial to evaluate the anti-aging effect of a day cream containing 0.6% by weight of an aqueous extract of Anigoxanthos plavidus . As a result of these tests, it was proved that the Anisoxanthos plavidus extract exhibits a contraction-remodeling effect on the skin by significantly reducing the depth of wrinkles for a long period of time. Angioxanthos plavidus extract exhibits an immediate contraction effect, which can be observed within minutes after application to the skin.

In connection with these results, the Anisoxanthos plavidus extract may therefore find application in the cosmetic field as an anti-aging agent for preventing or treating signs of skin aging. Angioxanthos plavidus extract may also find its application as an immediate-acting tightening agent to re-tighten the skin and improve skin appearance or skin retention for a short period of time.

Due to its action on the synthesis of several key proteins involved in the organization, cohesion and elasticity of the skin extracellular matrix, Angioxanthos flavidus extract is used for remodeling and remodeling of the extracellular matrix, especially the dermis in aged or aged skin. can be used to promote In general, angioxanthos plavidus extract can be used to treat or prevent skin sagging, improve skin elasticity, or treat or prevent skin wrinkles and fine lines. Thus, Anigoxanthos plavidus extract can be used as a tightening or lifting agent or as an agent for skin remodeling and restructuring. In particular, Angioxanthos plavidus extract is associated with loss of skin elasticity and/or changes in the extracellular matrix of the skin, for example, destruction of the extracellular matrix due to qualitative or quantitative changes in extracellular matrix components such as elastic fibers. It can be used as a cosmetic agent for treating or preventing non-pathological changes of the skin.

- Use of the Anisoxanthos plavidus extract according to the present invention

According to a first aspect, the present invention relates to non-therapeutic cosmetic use of an extract of Xanthos plavidus . Angioxanthos plavidus extract can be used as a cosmetic agent, especially as an anti-aging agent.

In the context of the present invention, an Anisoxanthos plavidus extract refers to an extract obtained from the aerial parts of Anisoxanthos plavidus , such as leaves, flowers, stems and combinations thereof. In certain embodiments, the Anisoxanthos plavidus extract is an extract obtained from the flowers and/or stems of Anisoxanthos plavidus . Angioxanthos plavidus extract can be obtained by any extraction technique. Preferably, the Anisoxanthos plavidus extract is prepared by extraction with a polar solvent. Generally, the polar solvent may be selected from water, alcohols, esters, ketones and mixtures thereof. As examples of esters, mention may be made of ethyl acetate, propyl acetate and butyl acetate. Alcohols include, but are not limited to, polyols such as methanol, ethanol, n-propanol, isopropanol, methanol and glycerol. As examples of ketones, mention may be made of acetone, cyclohexanone or methyl ethyl ketone.

In certain embodiments, the extraction solvent is selected from water, alcohols and aqueous-alcoholic mixtures such as water/ethanol, water/methanol or water/glycerol mixtures. In some specific embodiments, the extraction solvent is water.

Extraction with polar solvents can be performed by any method known to those skilled in the art, for example by decoction, infusion, maceration, percolation or leaching. can be performed For example, Angioxanthos plavidus extract can be obtained by thermal extraction in water or aqueous alcohol mixtures. A suitable heat extraction technique is decoction.

The term "decoction" means boiling the plant material in water for a time suitable to obtain efficient extraction.

The extraction step may be performed on the plant material of Anisoxanthos plavidus that has been subjected to one or more treatments such as freezing, drying, freeze drying, grinding or a combination thereof. In some embodiments, extraction is performed on fresh or dried plant material that has undergone a grinding step. At the end of the extraction, the extract may be recovered by any method capable of separating the plant material from the liquid phase, for example by centrifugation and/or filtration.

For example, angioxanthos plavidus extract,

- preparing plant material by drying and crushing the aerial parts, generally flowers and stems, of Angioxanthos plavidus ;

- extracting the plant material with a polar solvent, usually water or an aqueous-alcoholic mixture, preferably under high temperature conditions, for example by decoction, and

- preferably by a process comprising the step of removing the solid plant material by filtration.

The process for preparing the Angioxanthos plavidus extract may include one or more additional steps at the end of the extraction and/or filtration steps. The process may include, for example, formulating the extract by adding pH adjusters, stabilizers and/or cosmetically acceptable carriers or supports, concentrating the extract or drying the extract.

In further or alternative embodiments, the Anisoxanthos flavidus extract comprises flavonoids, polyphenols and carbohydrates. The flavonoids may correspond to 1% to 15%, for example 5% to 15% or 1% to 8%, naringin equivalent relative to the dry matter of the extract; The polyphenols may represent from 5% to 30%, for example from 5% to 25%, from 12% to 20%, from 10% to 30% or from 15% to 20% by weight of chlorogenic acid equivalents by weight relative to the dry matter of the extract, ; Sugars may represent from 35% to 65%, for example from 35% to 55%, from 40% to 60% or from 40% to 50% by weight of glucose equivalents relative to the dry matter of the extract. The content of polyphenols, flavonoids and sugars was determined by chlorogenic acid (Folin-Ciocalteu method), naringin (Zhishen et al. method, Food Chemistry, 1999, 64, 555-559) and glucose (Bachelier et al. method, Cah. OSTRUM.sec., Pudol Vo.IV, 3-1966), respectively, determined by colorimetry using standard substances obtained from

The Anisoxanthos plavidus extract obtained at the end of the extraction process comprises a dry matter content of from 0.1% to 40% by weight, preferably from 0.1% to 10% by weight, for example from 0.1% to 4% by weight. can do.

In certain embodiments, the Anisoxanthos plavidus extract comprises less than 1% by weight of ferulic acid on a dry matter basis of the extract. Preferably, the Anisoxanthos plavidus extract is free of ferulic acid.

In certain embodiments, the Anisoxanthos plavidus extract is characterized by one or more (e.g., two, three, four, five, or all) of the following characteristics:

- Anisoxanthos plavidus extract is obtained from the aerial parts of Anisoxanthos plavidus by extraction with polar solvents, preferably by decoction in water or in an aqueous-alcoholic mixture;

- the Anisoxanthos plavidus extract comprises a dry matter content of between 0.1% and 10% by weight;

- the flavonoids represent between 5% and 15%, for example between 7% and 13%, by weight of naringin equivalents relative to the dry matter of the extract;

- the sugars represent between 35% and 65%, for example between 40% and 60%, by weight of glucose equivalents relative to the dry matter of the extract;

- the polyphenols represent between 5% and 25%, for example between 10% and 20%, by weight of chlorogenic acid equivalents, relative to the dry matter of the extract; and/or

- Angioxanthos plavidus extract contains less than 1% by weight of ferulic acid in relation to the dry matter of the extract.

For use according to the present invention, the Anisoxanthos plavidus extract may be in liquid form, for example in water or in an aqueous-alcoholic mixture such as a water/glycerol mixture. In another embodiment, the Anisoxanthos plavidus extract may be in dry solid form, usually in powder form. The dry extract of Angioxanthos plavidus can be obtained by removing the extraction solvent by any known technique, for example spray drying, evaporation or freeze drying.

As a further example, the Anisoxanthos plavidus extract may be in an adsorbed form on a solid carrier such as maltodextrin.

In certain embodiments, the Anisoxanthos plavidus extract is a liquid extract in water or an aqueous-alcoholic mixture, preferably a water/glycerol mixture. The aqueous-alcoholic mixture may correspond to a water/alcohol volume ratio ranging from 1/9 to 9/1, preferably from 1/9 to 2/1.

For the purposes of the present invention, the term " anti-aging agent " is intended to mean a cosmetic active agent having biological activity capable of preventing or treating the signs of aging of the skin.

The expression “ preventing signs of skin aging ” means delaying or preventing the appearance of signs of skin aging.

The expression “ treating signs of skin aging ” means reducing, diminishing, attenuating, correcting or slowing down the development of signs of skin aging.

The expression “ signs of aging of the skin ” means non-determination of the visual appearance or mechanical properties of the skin due to aging, whether chronological (chronological aging) and/or photo-induced (photo-aging). Means a pathological change or transformation. Signs of skin aging thus refer to non-pathological changes or alterations in the appearance or mechanical properties of the skin.

Thus, signs of aging include thinning of the skin, especially of the epidermis, appearance of fine wrinkles, appearance of fine lines and/or wrinkles on the skin, including lips and eyelids, skin atrophy or sagging, loss of skin radiance, dull complexion, Pigmented spots on the skin, dark circles under the eyes, loss of skin density, loss of skin firmness, loss of skin tone, loss of skin contractility, loss of skin elasticity, change in the smooth appearance of the skin, increase in skin roughness and contouring of the face Changes in include, but are not limited to. For the purposes of the present invention, the terms “ skin ” and “ cutaneous ” refer to any part of human skin, such as the skin of the face, especially the lips and eyelids, the skin of the neck and the skin of the hands.

As suggested above, Anisanthos plavidus extract can promote the synthesis of various key proteins related to the structure of the extracellular matrix and skin elasticity.

Thus, the Angioxanthos plavidus extract can be used to improve, restore or prevent changes in the mechanical properties of the skin, in particular in the flexibility, elasticity, firmness or tonicity of the skin. For example, angioxanthos plavidus extract can be used to improve, restore or prevent aging skin or changes in the mechanical properties of aged skin.

The term " aging skin " refers to the skin of a man or woman over the age of 40 and refers to the loss of elasticity due to aging.

In a particular embodiment, the extract according to the present invention is used to treat or prevent signs of skin aging associated with changes in the mechanical properties of the skin, in particular loss of skin elasticity. Angioxanthos plavidus extract can thus be used as an agent for smoothing, swelling, lifting or relifting, restructuring or reshaping the skin. Angioxanthos plavidus extract can be used as an agent for immediate skin contraction.

The expression " agent for smoothing " means a cosmetic agent capable of reducing and/or correcting fine curves of the skin, including the lips, which are present in the form of wrinkles or fine lines.

The expression " agent for lifting or relifting " means a cosmetic agent capable of exerting a contracting action on the skin, thereby improving the appearance of the skin and making wrinkles less visible.

The expression “ agent for remodeling the skin ” means an agent capable of reducing the irregular appearance of the skin after loss of skin elasticity or sagging of the skin, especially in the contours of the face and neck.

The term " lifting agent " refers to a cosmetic agent capable of exerting a contracting effect on the skin to smooth out micro-curves of the skin.

The term " restructuring agent " is a cosmetic agent capable of preventing or treating changes in the extracellular matrix of the skin due to aging, such as loss or change of elastic fibers in the extracellular matrix or loss or change of collagen in the extracellular matrix. means

The expression "immediate tensioning agent or immediate-effect tensioning agent " refers to cosmetic products capable of exerting a contracting effect on the skin to increase skin elasticity and/or improve the appearance or retention of the skin. The effect is generally visible within a few minutes (e.g., 5 minutes) to 1 hour after application to the skin.

For example, angioxanthos plavidus extract may be used for the appearance of fine lines and/or wrinkles on the skin, especially on the lips or eyelids, atrophy on the skin, for example on the neck, sagging or sagging of the skin, loss of skin density. , loss of skin firmness, loss of skin tone, loss of skin elasticity, sagging of the skin, changes in the smooth appearance of the skin, increase in skin roughness, and changes in facial contours. Angioxanthos plavidus extract can be used to give mature or aged skin a more youthful appearance and/or to improve the complexion or radiance of aged or aged skin.

Angioxanthos plavidus extract can be used for cosmetic purposes to restructure or reconstruct the extracellular matrix of skin damaged due to aging. In particular, the Angioxanthos plavidus extract can be used for cosmetic purposes to stimulate the synthesis of proteins involved in the organization of the extracellular matrix of the skin. The expression " proteins involved in the organization of the extracellular matrix of the skin" refers to the expression or Means a protein involved in tissue. In some embodiments, the Anisoxanthos plavidus extract is used as a cosmetic agent to stimulate the synthesis of a protein selected from procollagen type I, collagen type XVII and tenascin-X in the skin, preferably aged skin. In an embodiment, the Anisoxanthos plavidus extract is used as a cosmetic agent to increase the contractility of dermal fibroblasts of aged skin or aged skin. In another embodiment, the Anisoxanthos plavidus extract is used to stimulate metabolism in the dermis.

In some embodiments, the Anisoxanthos plavidus extract is used in combination with an active agent that has a cosmetic effect. These agents are described below and include, among others, anti-wrinkle agents, anti-aging agents, antioxidants, moisturizing agents, lifting agents and firming agents.

In the use according to the present invention, the Anisoxanthos plavidus extract is applied to the skin. The extract is preferably present as a cosmetic agent in a composition for application to the skin. The composition is generally a cosmetic composition. Anisoxanthos plavidus extract generally corresponds to 0.0001% to 10% by weight, preferably 0.001% to 5% by weight, more preferably 0.005% to 3% by weight relative to the total weight of the cosmetic composition. do. For example, the Angioxanthos plavidus extract corresponds to 0.01% to 3.0% by weight, for example 0.6% by weight of the cosmetic composition. Angioxanthos plavidus extract can be added to a cosmetic composition in the form of a liquid extract, in the form of a dry extract, or in the form of a "precursor" composition as described later herein.

The cosmetic composition may contain one or more cosmetically acceptable excipients and optionally one or more additional active agents having a cosmetic effect. In general, the cosmetic composition,

- 0.0001% to 10% of anisoxanthos plavidus extract,

- 0% to 20% of one or more additional active agents, and

- from 70% to 99.9999% of one or more cosmetically acceptable excipients,

Percentage represents the weight percentage relative to the total weight of the cosmetic composition

In some embodiments, the cosmetic composition

- from 0.001% to 5% by weight of anisoxanthos flavidus extract,

- from 0.001% to 10% of one or more additional active agents, and

- from 85% to 99.998% of one or more cosmetically acceptable excipients.

In another embodiment, the cosmetic composition,

- from 0.01% to 5% by weight of anisoxanthos plavidus extract,

- from 0.01% to 10% of one or more additional active agents, and

- 85% to 99.98% of one or more cosmetically acceptable excipients.

Angioxanthos plavidus extract is preferably in liquid form.

In a further or alternative embodiment, the cosmetic composition comprises from 0.001% to 10%, for example from 0.005% to 1% or from 0.005% to 0.5% by weight of dry matter from the Xanthos plavidus extract. include

The expressions " active ingredient having a cosmetic effect ", " active agent having a cosmetic effect " or " cosmetic or active agent having a cosmetic effect " mean a compound capable of exerting at least one cosmetic effect on the skin or skin appendages. The term " cosmetic effect " is intended to modify and/or improve the appearance of the skin or mucous membranes such as lips, to protect the skin, etc. against external aggression (sun, wind, moisture, dryness, chemical products), or to prevent aging of the skin, etc. means a non-therapeutic effect aimed at preventing and/or correcting the phenomena associated with

Thus, in some embodiments, the Anisoxanthos plavidus extract may be present in a cosmetic composition that also includes additional active ingredients having a cosmetic effect. Active ingredients with cosmetic effects include vitamins, sun filters and sunscreens, anti-aging agents, anti-redness agents, antioxidants, firming agents, moisturizers, soothing agents, scrubbing or exfoliating agents, matting agents, sebum control agents, lightening actives, anti-spot agents, slimming agents , dehydrating agents, self-tanning actives, tanning accelerators, and combinations thereof. In particular, the cosmetic composition may include a plant extract such as tocopherol and/or linseed extract, a Vibrio exopolysaccharide extract , or a peptide such as trifluoroacetyl tripeptide-2.

Preferably, the cosmetic composition may include an active agent selected from the group consisting of an anti-wrinkle agent, an anti-redness agent, an antioxidant, a moisturizing active agent, a soothing agent, a sebum control agent, an anti-spot agent, a sun filter or sunscreen, and combinations thereof. More preferably, the additional active having a cosmetic effect is selected from anti-wrinkle agents, antioxidants, humectants, firming agents and combinations thereof.

As examples of humectants, urea, phydolic acid (PCA) and its derivatives, in particular arginine PCA, chitosan PCA, copper salt (copper PCA), magnesium salt (magnesium PCA), sodium salt (sodium PCA), or zinc salt, ethylhexyl PCA, calcium gluconate, hyaluronic acid and its salts and other glycosaminoglycans, fructose, glucose, isomaltose, lactose, trehalose, polydextrose, sucrose (sucrose), maltitol, mannitol, sorbitol , xylitol and other carbohydrates and derivatives, polyethylene glycols such as PEG-7, PEG-8, PEG-10, PEG-12 or PEG-14, glycerol, propylene glycol, butylene glycol, betaine, citrulline, collagen and derivatives thereof , histidine, silk hydrolysate, keratin hydrolysate or soybean hydrolysate, plant extracts rich in polysaccharides and/or polyphenols such as aloe extract, cornflower ( Centaurea cyanus ), and combinations thereof. can mention

Examples of anti-aging agents include ascorbic acid and its derivatives, such as magnesium ascorbyl phosphate, glycosaminoglycan and its derivatives, Cyathea polysaccharides, collagen, flaxseed ( Linum usitatissimum ) extract, Caprooil tetrapeptide-3 and trifluoroacetyl tripeptide-2, polygonum aviculare extract, brown algae, especially Ascophyllum nodosum extract , ferns, especially Cyathea cumingii extract. can do

Examples of sedatives include allantoin, aloe extract, birch (e.g. Betula alba ) extract, willow herb (e.g. Epilobium angustifolium ) extract, chestnut ( For example, Castanea sativa extract, cornflower (eg Centaurea cyanus ) extract, centella asiatica ( Centella asiatica ) extract, horsetail ( For example, Equisetum arvense ) extract, fennel (e.g., Foeniculum vulgare ) extract, cornflower (e.g., Hamamelis virginiana ) extract, ivy (eg Hedera helix ) extract, hibiscus sabdariffa extract, lily (eg Lilium candidum ) extract , mallow (eg , Malva sylvestris ( Malva sylvestris ) extract, lemon balm (eg Melissa officinalis ) extract, thimble flower (eg Scutellaria baicalensis ) extract, Mimosa (eg Mimosa tenuiflora ) extract, Potentilla (eg Potentilla erecta ) extract, oligosaccharide extract or oligosaccharide, eg flax, palmitoyl tripeptide- Peptides such as 8 and combinations thereof may be mentioned.

Examples of antioxidants include HMR (hydroxymethyl resorcinol), ascorbic acid and its derivatives, vitamin B9, histidine hydrochloride, or willow herb (e.g., Epilobium angustifolium ). angustifolium )) extract. Active ingredients of the vitamin type, which have antioxidant effects, are generally used in an amount of at least 1% by weight relative to the total weight of the cosmetic composition.

As examples of sebum control agents, mention may be made of flax lignan, rice flour, zinc gluconate, sarcosine, cinnamon extract, avocado extract and combinations thereof.

Examples of anti-redness agents include saponins, flavonoids, ruscogenin, esculoside , and extracts containing them, such as Ruscus extract and some essential oils, such as Mention may be made of lavender or rosemary.

As an example of an anti-spot agent, licorice ( Glycyrrhiza glabra ) extract, jackfruit ( Artocarpus heterophyllus ) extract, ground yellow ( R. occidentalis ( R. occidentalis )) , plant extracts belonging to the citrus family, resveratrol, peptides such as oligopeptide-68 and nonapeptide-1, kojic acid, magnesium ascorbyl phosphate and combinations thereof.

Cosmetically acceptable excipient(s) present in the cosmetic composition include diluents, dispersants, gelling agents, emollients, vectoring agents such as polycationic polymers or phospholipids, gums, resins, solvents, especially lower alcohols, especially ethanol, isopropanol, dipropylene glycol, butylene glycerol, propanediol, glycerol, sorbitol, and propylene glycol, modified and polymerized starch, fillers such as titanium dioxide or metal stearates, preservatives, essential oils, pearlescent agents, Dyes, odor absorbers, pH adjusters or neutralizers, lubricants, thickeners, surfactants including anionic, cationic, amphiphilic or nonionic surfactants, humectants, humectants, fragrances, organic or inorganic pigments such as iron oxides, oils of plant origin or fats, oils such as animal fats, petrolatum, synthetic oils such as silicone oil (cyclomethicone), fatty alcohol esters, fluoro oils, waxes, modified clays, bentones, metal salts of fatty acids, silica, polyethylene, mica, preservatives, antibacterial agents, minerals, carriers such as spring or floral water, and/or other substances commonly used in formulations in the cosmetic or pharmaceutical field.

Angioxanthos plavidus extract can be included in all types of cosmetic compositions. Preferably, it is a cosmetic composition having a form suitable for topical administration, in particular a form suitable for dermal application. The cosmetic composition is an oil-in-water (O / W) or water-in-oil (W / O) or multiple (triple: W / O / W or O / W / O) emulsion, nanoemulsion, especially having a droplet size of less than 100 nm It may be in the form of an O/W nanoemulsion, an aqueous gel or dispersion or an aqueous or aqueous-alcoholic solution of a powder. The cosmetic composition according to the present invention may be in the form of lotion, milk, cream, salve, foam, solution or ointment. More generally, the compositions according to the invention may also be in cosmetic form of any type. These are cosmetic care products or make-up or body hygiene products such as lotions, milks, serums, aqueous or oily gels, emulsions, creams, cream-gels, skin care waters, ointments, balms, foundations, sprays, eye It can be a shadow, a stick, a lipstick, a gloss, a foam, a deodorant, a nourishing mask, a shower gel, or an exfoliating or scrubbing product. As a non-limiting example, Anisoxanthos plavidus can be present as an anti-aging, re-lifting or reshaping agent in a day cream for aged skin or aged skin. Angioxanthos plavidus extract can be present as an anti-aging agent in rejuvenating serums for the face or neck. As a further example, the Anisoxanthos plavidus extract may be present as a smoothing or reshaping active in a care cream for the hands, neck or chest.

In certain embodiments, the Anisoxanthos plavidus extract is present in a cosmetic composition together with a vectoring agent. In some embodiments, the Anisoxanthos plavidus extract is formulated by a vectorization system. Vectorized systems can be micelles, liposomes including unilamellar or multilamellar liposomes, niosomes, ethosomes, layered systems, nanosomes, lipid or polymeric vesicles, natural or non-natural. It may be selected from the group consisting of micro- or nanoparticles of polymers, hydrogels. Preferably, the Anisoxanthos plavidus extract is encapsulated in a vectorized system selected from liposomes and layered systems. Specific examples of liposomes and layered systems for embodying the present invention are described, inter alia, in French Patent Application Nos. 1358589 and 1262303, among others.

A further object according to the present invention is to treat or prevent signs of skin aging in a subject, comprising administering to the subject a cosmetically effective amount of an extract of Anisoxanthos plavidus, preferably by a topical route. makeup method for Angioxanthos plavidus extract is generally applied to the skin to be treated, for example the face, neck or hands. Preferably, the Angioxanthos plavidus extract is present in the cosmetic composition as described above, and is therefore applied in the form of a cosmetic composition, for example, in the form of a cream. As noted above, signs of skin aging can be attributed to changes in the mechanical properties of the skin, particularly loss of skin elasticity. Preferably, the cosmetic method according to the present invention is characterized by the appearance of wrinkles and/or fine lines, the appearance of pigment spots on the skin, atrophy of the skin, sagging or sagging of the skin, loss of skin density, loss of skin firmness, loss of skin tension, It is intended to treat or prevent signs of skin aging selected from loss of skin elasticity, sagging of skin, change in smooth appearance of skin, increase in skin roughness, change in facial contour, and combinations thereof.

An object of the present invention is to exhibit an immediate contraction effect on the skin, preferably aging skin, comprising the step of administering to the subject an extract of Anigoxanthos plavidus in a cosmetically effective amount, preferably by a topical route. makeup method for Angioxanthos plavidus extract is generally applied to the skin to be treated, for example the face, neck or hands.

Another object according to the present invention is to reconstitute or restructure the extracellular matrix of the skin and/or to synthesize a protein selected from procollagen type I, collagen type XVII and tenascin-X in the skin, preferably aged skin. A cosmetic method for stimulating and/or increasing the contractile force of dermal fibroblasts of aging skin, said method comprising administering to a subject a cosmetically effective amount of an Anisoxanthos plavidus extract, preferably topically. Include steps. In another embodiment, the Anisoxanthos plavidus extract is used to stimulate metabolism in the dermis or to reconstitute or restructure the dermis.

In the cosmetic methods and uses according to the present invention, the dose to be administered and the frequency of administration of the Anigoxanthos plavidus extract vary depending on the desired cosmetic effect, the characteristics of the subjects, in particular their sex, age and skin type. Generally, the Anisoxanthos plavidus extract can be applied to the area to be treated once or twice a day, generally in the morning and/or evening, for several weeks or months, eg, for at least three months. The area to be treated is generally selected from the group consisting of hands, neck, face or facial parts such as around the eyes, around the lips, and in the nasolabial folds. Angioxanthos plavidus extract can be applied in the form of a cosmetic composition as described below. The cosmetic composition generally contains 0.001% to 10% by weight of the extract, for example, 0.01% to 5% by weight.

For example, in order to obtain an anti-aging effect on the face, a patient may apply a cosmetic composition to the face in a dosage of 1 g to 2 g of 0.1% by weight of anisoxanthos plavidus extract morning and evening.

The subject may be a female or male, generally 30 years of age or older, preferably 40 years of age or older or 45 years of age or older. For example, the subject may be a woman over 45 years of age.

- a further object according to the invention

According to a further aspect, an object of the present invention is also a cosmetic composition containing an Anisoxanthos plavidus extract. In general, the cosmetic composition,

- 0.0001% to 10% of anisoxanthos plavidus extract,

- 0% to 20% of one or more additional active agents, and

- from 70% to 99.9999% of one or more cosmetically acceptable excipients,

Percentage represents the weight percentage relative to the total weight of the cosmetic composition

In some embodiments, the cosmetic composition,

- from 0.001% to 5% by weight of anisoxanthos flavidus extract,

- from 0.001% to 10% of one or more additional active agents, and

- from 85% to 99.998% of one or more cosmetically acceptable excipients.

In another embodiment, the cosmetic composition,

- from 0.01% to 5% by weight of anisoxanthos plavidus extract,

- from 0.01% to 10% of one or more additional active agents, and

- 85% to 99.98% of one or more cosmetically acceptable excipients.

It goes without saying that additional active agents and pharmaceutically acceptable excipients are as described above. Likewise, the cosmetic composition may be of any type as described above.

In particular, the Anisoxanthos plavidus extract may be in the form of a liquid extract.

In a further or alternative embodiment, the cosmetic composition comprises from 0.001% to 10% by weight, for example from 0.005% to 1% by weight, dry matter of the Anigoxanthos plavidus extract.

According to a further aspect, an object of the present invention is a composition incorporated into a cosmetic composition. Thus, this composition corresponds to a "precursor" composition.

Preferably, this composition for the preparation of a cosmetic composition,

- from 10% to 50%, preferably from 20% to 40% of Anigoxanthos plavidus extract,

- 50% to 90%, preferably 60% to 80% of a carrier, preferably an aqueous solvent, an organic solvent, preferably a lower alcohol such as ethanol, propanediol, butylene glycol, glycerol or isopropanol, lipophilic a carrier selected from agents and mixtures thereof; and

- optionally, from 0.01% to 30% of additional cosmetically acceptable excipients, preferably pH adjusters, buffers, vectorizers, antioxidants, preservatives, stabilizers, thickeners, emulsifiers, hydrophilic or lipophilic gelling agents, perfumes , mineral oil or organic oil, and a further excipient selected from combinations thereof,

Percentages represent weight percentages relative to the total weight of the composition.

It goes without saying that the Anisoxanthos plavidus extract can be as described above. For example, an Anisoxanthos plavidus extract can be obtained from the aerial part of Anisoxanthos plavidus using a polar solvent. In some embodiments, the Anisoxanthos plavidus extract is a liquid extract, for example in water or in an aqueous-alcoholic mixture such as a water/glycerol mixture.

In a specific embodiment, the composition for the preparation of a cosmetic composition,

- 20% to 40% of an aqueous extract of Anisoxanthos plavidus , and

- contains 60% to 80% glycerol,

Percentages represent weight percentages relative to the total weight of the composition.

A "precursor" composition may also include a buffering agent or a pH adjusting agent such as citric acid. Generally, a “precursor” composition according to the present invention has a pH between 3.0 and 4.5, usually around 3.9.

In a further or alternative embodiment, the “precursor” composition comprises from 0.01% to 10%, such as from 0.05% to 5%, or from 0.1% to 1% by weight of an Anisoxanthos plavidus extract. Contains dry matter.

An object of the present invention is also the use of said "precursor" compositions for the manufacture of cosmetic compositions. It goes without saying that the final cosmetic or pharmaceutical composition is particularly suitable for the practice of any one of the cosmetic uses according to the present invention and may have any of the above-mentioned characteristics.

The "precursor" composition is from 0.001% to 50% by weight, preferably from 0.01% to 10% by weight, for example from 0.5% to 5% by weight, or 1.5% by weight, relative to the total weight of the final cosmetic composition or pharmaceutical composition. % to 2.5% by weight of the cosmetic composition.

The cosmetic composition can be obtained by mixing the “precursor” composition according to the present invention with one or more excipients and/or one or more additional active agents having a cosmetic effect in terms of cosmetics.

A further object according to the present invention is a process for preparing the cosmetic composition according to the present invention, which is not a process, wherein the Xanthos plavidus extract or "precursor" composition has one or more cosmetically acceptable excipients and/or cosmetic effects. mixing with one or more additional active agents.

An object of the present invention is also a method for preparing a "precursor" composition according to the present invention, the method comprising:

a) supplying a plant substrate of Angioxanthos plavidus , preferably from the aerial parts of Angioxanthos plavidus, by drying and grinding;

b) extraction of the plant substrate using a polar solvent to obtain a liquid extract followed by filtration;

c) adding a cosmetically acceptable carrier to the liquid extract obtained by filtration;

d) optionally adding cosmetically acceptable excipients to the filtrate, and

e) optionally, adjusting the pH;

Steps c), d) and e) may be performed in any order.

In some embodiments, the cosmetically acceptable carrier includes glycerol and/or water.

The extraction step can be carried out as described above, for example using water or an aqueous-alcoholic mixture as the polar solvent.

Other aspects and advantages of the present invention will appear on reading the following examples, which are to be regarded as illustrative and in no way limiting.

Example

The following examples were carried out using a composition, hereinafter referred to as "extract", said "extract" composition comprising:

- 30% by weight of an aqueous extract of Anisoxanthos plavidus flowers or stems containing about 1.6% by weight of solids, and

- 70% by weight of glycerol.

For the purposes of the present invention, this is a "precursor" composition.

Example 1: Synthesis of Type I Procollagen by Human Dermal Fibroblasts Angioxanthos Flavidus Effect of Extract

- background

Collagen constitutes a family of proteins that are very involved in the mechanical properties of skin. Procollagen type I is a precursor to type I collagen, a major component of connective tissue. As we age, the collagen content in the skin decreases, resulting in wrinkles and fine lines forming on the skin.

The purpose of this study was not to elucidate the effect of Xanthos plavidus extract on the synthesis of procollagen by human dermal fibroblasts.

- Protocol

"Extract" compositions were tested at concentrations of 0.1% and 0.05% (v/v). TGF-β was selected as a positive control and tested at 10 ng/ml. Negative controls correspond to cells not treated with the "extract" composition or TGF-β

Fibroblasts were isolated from human dermis and cultured in specific DMEM medium supplemented with 10% fetal bovine serum, 1% antibiotics (penicillin/streptomycin) and 1% L-glutamine at 37°C, 5% CO ₂ and 95°C. % humidity was maintained.

2.5 x 10 ⁴ cells were seeded per microplate in complete DMEM medium. Cells were left in the incubator for 24 hours. The culture medium was then replaced with serum-free DMEM. The microplates were left in the incubator for an additional 24 hours to reach cell arrest.

The "extract" composition or TGF-β was added at the desired concentration. The microplate was incubated for an additional 24 hours. At the end of the culture, the supernatant was removed and analyzed by ELISA (MK101Z, Ozyme) to quantify type I procollagen.

- result

The obtained results are shown in FIG. 1 . Figure 1 shows the activation rate of type I procollagen synthesis in treated cells compared to untreated cells.

Fibroblasts treated with the “extract” composition or TGF-β produced higher amounts of procollagen type I than untreated cells. The effect of the "extract" composition on type I collagen synthesis is dose dependent. Angioxanthos plavidus extract stimulates type I procollagen in dermal fibroblasts.

Example 2: Synthesis of Tenascin-X by human skin fibroblasts Angioxanthos Flavidus Effect of Extract

Tenascin-X is a protein that regulates both the structure and stability of elastic fibers of the extracellular matrix (ECM). It is also involved in the organization of collagen fibers. Tenascin-X plays an important role in the strength and elasticity of connective tissue. A genetic deficiency of Tenascin-X can cause an inherited disease known as Ehlers-Danlos syndrome.

The purpose of this study was to determine the effect of the "extract" composition on tenascin-X synthesis by human dermal fibroblasts.

- background

The "extract" composition was tested at 0.2% and 0.1% (v/v). 10% fetal bovine serum was used as a positive control. Negative control corresponds to cells not treated with extract or fetal bovine serum.

Fibroblasts were isolated from human abdominal dermis and cultured in specific DMEM medium supplemented with 10% fetal bovine serum, 1% antibiotics (penicillin/streptomycin) and 1% L-glutamine at 37°C, 5% CO ₂ and 95°C. % humidity was maintained.

2.5 x 10 ⁴ cells were seeded per microplate in complete DMEM medium. Cells were left in the incubator for 24 hours. The culture medium was then replaced with serum-free DMEM. The microplates were left in the incubator for an additional 24 hours to reach cell arrest.

The "extract" composition or fetal bovine serum was added at the desired concentration. After addition, the cells were further cultured for 24 hours. At the end of the culture, the supernatant was removed and analyzed by ELISA to quantify Tenascin-X. At the same time, proteins contained in cell lysates at the end of culture were quantified by the Bradford method.

- result

The obtained results were subjected to Student's t-test when the p value was less than 0.05. The results are shown in FIG. 2 . Figure 2 shows the amount of Tenascin-X present in the medium, expressed as a percentage of the total amount of protein, for each experimental condition. It should be noted that the "extract" composition dose-dependently increased the amount of Tenascin-X compared to the negative control (untreated cells). In other words, Anisoxanthos plavidus extract stimulates Tenascin-X in human fibroblasts.

Example 3: Synthesis of Type XVII Collagen by Human Keratinocytes Angioxanthos Flavidus Effect of Extract

Type XVII collagen is a transmembrane protein that plays an essential role in maintaining the bond between intracellular and extracellular components involved in epidermal adhesion.

The purpose of this study was to determine the effect of the "extract" composition on type XVII collagen synthesis by human keratinocytes.

- Protocol

The "extract" composition was tested at a concentration of 0.4% (v/v). TGF-β was selected as a positive control and tested at 10 ng/ml. Negative controls correspond to cells not treated with the extract or TGF-β.

Keratinocytes were isolated from a human epithelial cell line and incubated at 37°C, 5% CO ₂ and 95% DMEM supplemented with 10% fetal bovine serum, 1% antibiotics (penicillin/streptomycin) and 1% L-glutamine. Humidity was maintained.

5 x 10 ⁴ cells were seeded per microplate in complete DMEM medium and left in the incubator for 24 hours. Then, the culture medium was replaced with DMEM containing 1% of fetal bovine serum, 1% of antibiotics and 1% of L-glutamine. The microplates were incubated for an additional 24 hours in an incubator.

At the end of the incubation, the "extract" composition or TGF-β was added at the desired concentration. The microplate was incubated for an additional 24 hours.

Type XVII collagen was detected using an anti-type XVII collagen antibody at a dilution of 1/100 and quantified by immunolabeling, expressed as a secondary antibody bound to fluorescein. Fluorescence intensity corresponds to the expression level of type XVII collagen. Images were processed using Image J software.

- result

Fluorescence measurements were normalized to the measurements of fluorescence intensity measured for negative controls. A +27% increase in fluorescence intensity was observed for cells cultured with the “extract” composition, showing an increase in type XVII collagen expression compared to untreated cells. Incubation with TGF-β (positive control) resulted in a 257% increase in fluorescence signal compared to the negative control.

In summary, Anisoxanthos plavidus extract stimulates type XVII collagen synthesis by human keratinocytes.

Example 4: Evaluation of contractile effect of extracts: Quantification of contractile force produced by fibroblasts from the bottom of wrinkles and quantification of α-SM actin

Using the GlasBox ^Plus™ system, it is possible to quantify the contractile effect of cosmetic actives on skin "equivalents" obtained from fibroblasts from the bottom of a wrinkle. Fibroblasts from the bottom of the fold develop less contractile force than adjacent unwrinkled skin. In addition, immunolabeling of α-SM actin can be performed in skin equivalents after measuring contractility in the GlasBox ^Plus™ system. The purpose of this study was to demonstrate the effects of the extract on fibroblast contractility and the synthesis of α-SM actin involved in cell plasticity, migration and motility.

- Protocol

- Fibroblast culture

A 2 mm diameter biopsy was taken from a wrinkle removal surgery (surgery waste: 63 year old female) at the level of unwrinkled skin and within the wrinkles.

Fibroblasts were extracted from healthy skin (HF) and wrinkle bottom (BW) using a conventional explantation technique. Explants were humidified at 37°C in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum (FCS), 40 mg/l gentamicin and 2 mg/l fungizone. and cultured in the presence of 5% CO ₂ . Slowly (over about 10 days), fibroblasts migrated out of the explants. The culture medium was changed twice a week. When fibroblasts are present in sufficient quantity, they are isolated under the action of trypsin-EDTA, subcultured and expanded under the same culture conditions.

- GlasBox ^PlusTM system and contractile force measurement

Skin equivalents are grown in Petri dishes consisting of eight rectangular cells. Two flexible slides are placed on each one, the bottom of which consists of a grid to which the skin equivalent is attached when polymerized. The skin equivalent grows between the two slides to form a rectangle with a slight contraction in the center. In conventional mechanics, this shape is referred to as a "diabolo" shape.

The slide distorts under the influence of the contractile force generated by the fibroblasts. Their distortion is measured through an optical fiber. This distortion is proportional to the force generated within the skin equivalent. Measurements are performed in real time using a PC acquisition card and appropriate software.

At confluency, monolayer cultured fibroblasts were trypsinized and then counted using a Malassez slide to obtain a cell suspension of 8×10 ⁵ cells/ml. 6x volume of 1.76X medium (DMEMc, NaHCO ₃ , NaOH, antibiotics, FCS), 1x volume of rat tail type I collagen (2mg/ml) and 1x volume of cell suspension (8 x 10 ⁵ cells/ml) was mixed to prepare a skin equivalent production medium.

The mixture was poured into the rectangular cell of the GlasBox ^Plus™ . The gel formed within minutes at 37°C. Various culture media containing the "extract" composition (final concentrations in the medium: 0.1% and 0.2% (V/V)) or TGF-β1 (positive control) or culture medium without the composition (negative control) were added and isotonic Isometric force was measured for 24 hours.

- Labeling of α-SM actin

Immunolabeling of α-SM actin fibers was performed using an anti-α-SM actin mouse monoclonal antibody. An anti-mouse secondary antibody coupled to FITC enables immunolabeling and screening. Nuclei were stained with Hoechst dye. Observations were performed by confocal microscopy at 488 nm and two-photon excitation microscopy at 705 nm.

Skin equivalent pieces were obtained using a 5 mm diameter biopsy punch. Skin equivalents were permeabilized by placing them in cold acetone. After washing with PBS, an aqueous hydrogen peroxide solution was added to remove endogenous peroxidase. After washing with PBS, the skin equivalent was placed in a glycine solution to remove background noise due to the aldehyde group of the fixative. Then, the skin equivalent was placed in a blocking solution capable of blocking non-specific binding due to the secondary antibody. Tissues were left in primary antibody overnight. After washing with PBS, secondary antibodies were added. After incubation for 1 hour and washing with PBS, nuclei were stained with Hoehst dye for 10 minutes.

After washing with PBS, skin equivalents were mounted between slides and cover slips by Eukitt. The slides were viewed and photographed under a confocal microscope (Zeiss LSM510 NLO) at x40 magnification.

- Statistical analysis

For the study of the contractile force, a two-way ANOVA was performed and, if necessary, a Fisher test was performed. Significance was maintained only when p<0.05. Area under the curve (AUC) was calculated as slope (shrinkage) and maximum shrinkage, all of which were calculated with GraphPad Prism® 5 software. One-way analysis of variance was performed on these data, and Fisher's test was performed when necessary. Significance was maintained only when p<0.05.

- result

The contractile force produced by fibroblasts at the bottom of the wrinkles (BW) is significantly smaller than that produced by healthy fibroblasts (HF). AUC (area under the curve representing contractile force over time) and maximal contraction were observed to be significantly reduced for BW fibroblasts compared to HF fibroblasts (data not shown). Culturing of fibroblasts with TGF-β1 induces a significant increase in AUC and maximal contractile force in both BW and HF fibroblasts.

Cultivation using the "extract" composition greatly increases the contractile force of the BW fibroblast population; Maximal contractile force and AUC are greatly increased. These results are illustrated in Figures 3A, 3B and 3C. The "extract" composition also increased contractility overall in HF fibroblasts, but the observed increase was less than that obtained in BW fibroblasts. Immunolabeling experiments demonstrate that the "extract" composition stimulates the expression of α-SM actin by BW fibroblasts (see Figure 4).

Angioxanthos plavidus extract significantly and distinctly stimulates the contractile force of fibroblasts from wrinkled skin. Thus, it exerts a contraction and lifting effect on aging skin or aged skin.

Example 5: Evaluation of the immediate physical contraction effect of the extract

Using the GlasBox ^Plus™ system, it was also possible to quantify the immediate contractile effect of the extract on its acellular skin equivalent by measuring the contractile force produced.

- Protocol

- GlasBox ^PlusTM system and contractile force measurement

The acellular skin equivalent polymerizes in a petri dish composed of eight rectangular cells. Two flexible slides are placed on each one, the bottom of which consists of a grid to which the acellular skin equivalent is attached when polymerized. Polymerization of the collagen gel, with or without the extract, creates a force that causes distortion of the slide. This distortion is measured through an optical fiber. This is proportional to the force generated within the non-cellular skin equivalent.

A skin equivalent production medium was prepared by mixing 6 volumes of 1.76X medium (DMEMc, NaHCO ₃ , NaOH, antibiotics, FCS), 3 volumes of rat tail type I collagen (2 mg/ml) and 1 volume of DMEMc. .

The mixture was poured into the rectangular cell of the GlasBox ^Plus™ . The gel formed within minutes at 37°C. Various culture media containing the "extract" composition (final concentration in the medium: 0.2% (V/V)) or culture medium without the composition (negative control) were added and isotonicity was measured for 24 hours.

- Statistical analysis

Two-way analysis of variance was performed for the study of contractile force, and Fisher's test (p <0.05) was performed when necessary.

Area under the curve (AUC), slope (shrinkage) and maximum shrinkage were calculated using GraphPad Prism®5 software. One-way analysis of variance was performed on these data, and where necessary, Fisher's test (p < 0.05) was performed.

- result

The contractile force is significantly increased by 0.2% compared to the control in the presence of the "extract". This increase is significant at 1 hour and 30 minutes after the start of the measurement and increases up to at least 11 hours in the presence of 0.2% extract (Figs. 5A and 5B). It was also observed that the AUC (area under the curve representing contractile force over time) and maximal contraction were significantly increased for the group treated with the extract according to the present invention compared to the control group not treated with the extract.

Angioxanthos plavidus extract significantly and distinctly stimulates the contractile force of the acellular skin equivalent. The Anigoxanthos plavidus extract according to the present invention thus exerts physical effects of immediate, astringent, shrinking and lifting, and thus can be used to exert an immediate tightening effect on aging skin.

Example 6: Examples of cosmetic compositions containing extracts

-An anti-aging cream comprising 2% of the "Extract" composition (Cream B )

The anti-aging properties of Cream B were evaluated in the clinical trial presented in Example 7.

-An anti-aging serum containing 2% of the "extract" composition

- anti-aging cream containing 2% of the "extract" composition

Example 7: Clinical evaluation of the anti-wrinkle effect of extracts

Cream tested :

- Cream B: contains 2% of the "Extract" composition (see Example 5).

- Cream A: A placebo cream having the same composition as Cream B except that 2% of the "extract" composition was replaced with 2% deionized water.

- Protocol

Testing for the use of Cream A product (placebo cream) was conducted in comparison to Cream B product containing 2% of the "extract" composition according to the following protocol:

Forty-two volunteers were blindly included in this study and classified into two groups according to a randomization list established by a dedicated program:

- Group A receiving Cream A: 21 women aged 45 to 69 years old (average age: 56.4 years) with all types of skin, of which 57% were sensitive skin, and skin phototypes II to IV.

- Group B receiving Cream B: 21 women (average age: 60.8 years), 46 to 77 years old, with all types of skin, of which 48% were sensitive skin, and skin phototypes II to IV.

The product was first applied to volunteers in the laboratory. Subsequent applications were administered voluntarily at the volunteer's home twice a day. T+1 hour after first application: After removing the product, the tolerance of the product was evaluated by a dermatologist's skin parameter evaluation for the affected area. Day T+7: Tolerance of the product was evaluated by evaluation of skin parameters (telephone contact) for the affected area after 7 days of use. Day T+28: Tolerance evaluation of the product was performed by a dermatologist according to the evaluation of skin parameters after 28 days of testing. Self-evaluation of product quality was conducted by volunteers through a dedicated questionnaire.

In addition, the efficacy of the product was evaluated for the average depth of crow's feet.

Efficacy measurements were taken at T0, T1h (ie 1 hour after application) and T28 (ie 28 days after application of the tested product). As a result of the measurement at 1 h, it was possible to evaluate the immediate contraction effect of the extract. As a result of the measurement at T28, it was possible to evaluate the anti-aging effect of the extract, especially the long-term anti-wrinkle effect. Evaluation of wrinkle depth was performed by quantifying the curvature of the skin surface using the LifeViz Micro™ system (Quantificare). LifeViz Micro™ is a non-contact measurement method based on the projection of structured light onto a given skin area.

- result

Cream B was well tolerated by the patients during the trial.

Skin treated with cream containing Angioxanthos plavidus extract showed a significant 10% reduction in wrinkle depth after 28 days of application compared to skin treated with Cream A (placebo cream) ( FIG. 6 ). A reduction in wrinkle depth of 5% can be seen 1 hour after application of Cream B, which clearly demonstrates the immediate tightening effect of the extract according to the present invention (Fig. 6).

Claims (13)

Skin, comprising Anigozanthos flavidus extract as a cosmetic agent for smoothing, lifting, relifting, restructuring or remodeling the skin An anti-aging cosmetic composition for improving, restoring or preventing change in mechanical properties of
According to claim 1,
A cosmetic composition in which an anisoxanthos plavidus extract is present as an agent for immediate tensioning of the skin.
According to claim 1,
The cosmetic composition Atrophy or sagging of the skin, loss of skin density, loss of skin firmness, loss of skin tension, loss of skin elasticity, loss of skin contractility, loss of skin retention, skin sagging, changes in the smooth appearance of the skin and changes in the contours of the face For alleviating or preventing the sign (s) of skin aging due to changes in the mechanical properties of the skin selected from the group consisting of, a cosmetic composition.
According to claim 1,
Anisoxanthos plavidus extract is obtained from the aerial part of Anisoxanthos plavidus selected from flowers, leaves, stems and combinations thereof, a cosmetic composition.
According to claim 4,
Anisoxanthos plavidus extract is obtained by extraction with a polar solvent, a cosmetic composition.
According to any one of claims 1 to 5,
Cosmetic compositions include vitamins, sun filters and sunscreens, anti-aging or anti-wrinkle agents, antioxidants, lifting agents, firming agents, anti-spot agents, anti-redness agents, and slimming agents. agent), dehydrating agent, humectant, emollient, scrubbing or exfoliating agent, matting agent, sebum control agent, lightening active, self-tanning active, tanning accelerator and A cosmetic composition that also includes at least one additional cosmetic agent selected from the group consisting of combinations thereof.
According to any one of claims 1 to 5,
The cosmetic composition may be an aqueous solution, an aqueous-alcoholic solution, an oil-in-water (O/W) or water-in-oil (W/O) or multiple (triple: W/O/W or O/W/O) emulsion, a nanoemulsion, O/W nanoemulsions with droplet sizes of less than 100 nm, aqueous gels or dispersions of the fatty phase in the aqueous phase with spherules, suspensions, liposomal suspensions, powders, lotions, milks, creams, plasters, gels, foams and Which is selected from the group consisting of ointment, a cosmetic composition.
According to any one of claims 1 to 5,
Cosmetic compositions include cosmetics, make-up products, body hygiene products, lotions, milk, serums, aqueous or oily gels, emulsions, creams, cream-gels, skin care waters, ointments, balms, foundations, sprays, eye shadows, and sticks. , a cosmetic composition selected from lipsticks, glosses, lip balms, foams, deodorants, nourishing masks, shower gels, and peeling or scrubbing products.
According to any one of claims 1 to 5,
A cosmetic composition for improving, restoring or preventing loss of skin elasticity or tonicity.
According to any one of claims 1 to 5,
A cosmetic composition intended for use on mature skin or older skin.
According to any one of claims 1 to 5,
The extract of Anisoxanthos plavidus is a cosmetic agent for stimulating the synthesis of proteins selected from procollagen I, collagen XVII, and Tenascin-X in the skin. , Or, which is used as a cosmetic agent for increasing the contractility of dermal fibroblasts of aged skin or aged skin, a cosmetic composition.
A process for producing the cosmetic composition according to any one of claims 1 to 5,
The process is not a cosmetic comprising mixing a composition comprising 20% to 40% by weight of an aqueous extract of Xanthos plavidus and 60% to 80% by weight of glycerol with one or more cosmetically acceptable excipients Process for preparing the composition.
13. The process for preparing a cosmetic composition according to claim 12, wherein at least one additional active agent having a cosmetic effect is mixed with the composition.

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